Date of Award

6-2015

Degree Name

MS in Biological Sciences

Department

Biological Sciences

Advisor

Marie Yeung, Ph.D.

Abstract

Vibrio parahaemolyticus is a food-borne pathogen commonly associated with the consumption of raw or undercooked seafood resulting in primary infections of the human gastrointestinal tract. It is estimated to cause about 4500 illnesses each year in the United States. However, infection from this food-borne pathogen can be avoided if this organism is detected in the implicated food, prior to consumption. Current standard methods of detecting this organism are dependent on the culturability of the bacteria. Detection based on an organism’s culturability may be problematic as V. parahaemolyticus has been known to exist in a viable but nonculturable (VBNC) state. Bacteria in the VBNC state are characterized by low levels of metabolic activity and the inability to be cultured by standard laboratory practices. When bacteria enter the VBNC state, their gene expression profile may be different than the culturable counterpart. We were interested in comparing the expression of two virulence-associated genes between VBNC and culturable cells of V. parahaemolyticus. V. parahaemolyticus RIMD2210633 was incubated at 4°C in modified Morita mineral salt solution supplemented with 0.5% NaCl (MMS) or trypticase soy broth supplemented with 2% NaCl (TSBS), which represented nutrient poor and rich conditions, respectively. The number of VBNC and culturable cells were determined by standard plate count and fluorescence microscopy. The expression levels of virulence-associated genes tdh2 and escU, were measured relative to the housekeeping gene, pvsA, by qRT-PCR. Nutrient availability and temperatures exerted variable effects on the virulence gene expression. It is possible that VBNC V. parahaemolyticus cells may retain their pathogenicity potential.

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